Isoquinoline derivatives

ABSTRACT

Isoquinoline derivatives of the general formula (I) in which X, Y, Z, R 1 , R 2  and n are as defined in Patent Claim ( 1 ), and physiologically acceptable salts or solvates thereof, are integrin inhibitors and can be employed for combating thromboses, cardiac infarction, coronary heart diseases, arteriosclerosis, inflammation, tumors, osteoporosis, infections and restenosis after angioplasty or in pathological processes which are maintained or propagated by angiogenesis.

The invention relates to compounds of the general formula I

in which

-   -   X is H, —C(═NR³)—NHR⁴ or Het,

-   -   Y is —(CH2)m-,    -   Z is NH or CH₂,    -   R¹ and R⁵ are each, independently of one another, H, A, OH, OA,        arylalkyl, Hal, —CO—A, CN, NO₂, NHR³, COOA, COOH, SO₂A, CF₃ or        OCF₃,    -   R² is in each case, independently of the others, H or A,    -   R³ and R⁴ are each, independently of one another, H, A, —CO—A,        NO₂ or CN.    -   A is alkyl having 1–6 carbon atoms,    -   m is 0, 1, 2, 3, 4, 5 or 6,    -   n and p are, independently of one another, 1, 2 or 3,    -   and physiologically acceptable derivatives thereof, in        particular salts and solvates thereof.

The invention had the object of finding novel compounds having valuableproperties, in particular those which are used for the preparation ofmedicaments.

It has been found that the compounds of the formula I and salts thereofhave very valuable pharmacological properties and are well tolerated. Inparticular, they act as integrin inhibitors, inhibiting, in particular,the interactions of the αv, β3, β5 or β6 integrin receptors withligands, such as, for example, the binding of fibrinogen to the integrinreceptor.

Integrins belong to the family of heterodimeric class I transmembranereceptors, which play an important role in numerous cell-matrix andcell-cell adhesion processes (Tuckwell et al., 1996, Symp. Soc. Exp.Biol. 47). They can be divided roughly into three classes: the β1integrins, which are receptors for the extracellular matrix, the β2integrins, which can be activated on leukocytes and are triggered duringinflammatory processes, and the αv integrins which influence the cellresponse during wound-healing and other pathological processes (Marshalland Hart, 1996, Semin. Cancer Biol. 7, 191). The relative affinity andspecificity for ligand binding is determined by the combination of thevarious α and β sub-units.

The compounds according to the invention exhibit particulareffectiveness in the case of integrins αvβ1, αvβ3, αvβ5, αIIbβ3 as wellas αvβ6 and αvβ8, preferably of αvβ3, αvβ5 and αvβ6, as well as αIIbβ3.

αvβ6 is a relatively rare integrin (Busk et al., J. Biol. Chem. 1992,267(9), 5790), which is formed to an increased extent during repairprocesses in epithelial tissue and which preferably binds the naturalmatrix molecules fibronectin and tenascin (Wang et al., Am. J. Respir.Cell Mol. Biol. 1996, 15(5), 664). The physiological and pathologicalfunctions of αvβ6 are not yet known precisely, but it is assumed thatthis integrin plays an important role in physiological processes anddiseases (for example inflammation, wound healing, tumours) in whichepithelial cells are involved. Thus, αvβ6 is expressed on keratinocytesin wounds (Haapasalmi et al., J. Invest. Dermatol. 1996, 106(1), 42),from which it can be assumed that, besides wound-healing processes andinflammation, other pathological events of the skin, such as, forexample, psoriasis, bullate pemphigus, dermatitis and erythema and alsocystic fibrosis, endometriosis, liver cirrhosis and periodontitis, canalso be influenced by agonists or antagonists of the said integrin.Furthermore, αvβ6 plays a role in the respiratory tract epithelium(Weinacker et al., Am. J. Respir. Cell Mol. Biol. 1995, 12(5), 547), andconsequently corresponding agonists/antagonists of this integrin couldsuccessfully be employed in respiratory tract diseases, such asbronchitis, asthma, lung fibrosis and respiratory tract tumours.Finally, it is known that αvβ6 also plays a role in the intestinalepithelium, which means that the corresponding integrinagonists/antagonists could be used in the treatment of inflammation,tumours and wounds of the gastric/intestinal tract.

It has been found that the compounds of the formula I according to theinvention and salts thereof exert, as soluble molecules, an action oncells which carry the said receptor or, if they are bonded to surfaces,are artificial ligands for αvβ6-mediated cell adhesion. In particular,they act as αvβ6 integrin inhibitors, inhibiting, in particular, theinteractions of the receptor with other ligands, such as, for example,the binding of fibronectin.

The compounds according to the invention are, in particular, potentinhibitors of the vitronectin receptor αvβ3 and/or potent inhibitors ofthe αvβ6 receptor.

αvβ3 integrin is expressed on a number of cells, for example endothelialcells, cells of smooth vascular muscles, for example of the aorta, cellsfor breaking down bone matrix (osteoclasts) or tumour cells.

The action of the compounds according to the invention can bedemonstrated, for example, by the method described by J. W. Smith et al.in J. Biol. Chem. 1990, 265, 12267–12271.

B. Felding-Habermann and D. A. Cheresh in Curr. Opin. Cell. Biol. 1993,5, 864, describe the importance of the integrins as adhesion receptorsfor a very wide variety of phenomena and syndromes, especially withrelation to the vitronectin receptor αvβ3.

The dependence of the occurence of angiogenesis on the interactionbetween vascular integrins and extracellular matrix proteins has beendescribed by P. C. Brooks, R. A. Clark and D. A. Cheresh in Science1994, 264, 569–571.

The possibility of inhibiting this interaction and thus initiatingapoptosis (programmed cell death) of angiogenic vascular cells by acyclic peptide has been described by P. C. Brooks, A. M. Montgomery, M.Rosenfeld, R. A. Reisfeld, T. Hu, G. Klier and D. A. Cheresh in Cell1994, 79,1157–1164. This described, for example, αvβ3 antagonists orantibodies against αvβ3 which cause shrinkage of tumours due to theinitiation of apoptosis.

The experimental evidence that the compounds according to the inventionalso prevent the adhesion of living cells to the corresponding matrixproteins and accordingly also prevent the adhesion of tumour cells tomatrix proteins can be provided in a cell adhesion test analogously tothe method of F. Mitjans et al., J. Cell Science 1995, 108, 2825–2838.

P. C. Brooks et al. in J. Clin. Invest. 1995, 96, 1815–1822, describeα_(v)β₃ antagonists for combating cancer and for the treatment oftumour-induced angiogenic diseases.

The compounds are able to inhibit the binding of metal proteinases tointegrins and thus prevent the cells from being able to utilise theenzymatic activity of the proteinase. An example is the possibility ofinhibiting the binding of MMP-2-(matrix metalloproteinase 2-) to thevitronectin receptor αvβ3 by a cyclo-RGD peptide, as described in P. C.Brooks et al., Cell 1996, 85, 683–693.

The compounds of the formula I according to the invention can thereforebe employed as medicament active ingredients, in particular for thetreatment of tumour diseases, osteoporosis, osteolytic diseases and forthe suppression of angiogenesis.

Compounds of the formula I which block the interaction of integrinreceptors and ligands, such as, for example, of fibrinogen to thefibrinogen receptor (glycoprotein IIb/IIIa), prevent, as GPIIb/IIIaantagonists, the spread of tumour cells by metastasis. This is confirmedby the following observations:

The spread of tumour cells from a local tumour into the vascular systemtakes place through the formation of microaggregates (microthrombi)through the interaction of the tumour cells with blood platelets. Thetumour cells are screened by the protection in the microaggregate andare not recognised by the cells of the immune system. Themicroaggregates can attach themselves to vascular walls, simplifyingfurther penetration of tumour cells into the tissue. Since the formationof the microthrombi is promoted by fibrinogen binding to the fibrinogenreceptors on activated blood platelets, the GPIIb/IIIa antagonists canbe regarded as effective metastasis inhibitors.

Besides the binding of fibrinogen, fibronectin and von Willebrand factorto the fibrinogen receptor of the blood platelets, compounds of theformula I also inhibit the binding of further adhesive proteins, such asvitronectin, collagen and laminin, to the corresponding receptors on thesurface of various types of cell. In particular, they prevent theformation of blood-platelet thrombi and can therefore be employed forthe treatment of thromboses, apoplexia, cardiac infarction, inflammationand arteriosclerosis.

The thrombocyte aggregation-inhibiting action can be demonstrated invitro by the method of Born (Nature 1962, 4832, 927–929).

A measure of the take-up of a medicament active ingredient into anorganism is its bioavailability.

If the medicament active ingredient is administered to the organismintravenously in the form of an injection solution, its absolutebioavailability, i.e. the proportion of the pharmaceutical species whichis unchanged in the systemic blood, i.e. enters the general circulation,is 100%.

On oral administration of a therapeutic active ingredient, the activeingredient is generally present in the formulation in the form of asolid and must therefore first dissolve in order that it can overcomethe entry barriers, for example the gastrointestinal tract, the oralmucous membrane, nasal membranes or the skin, in particular the stratumcorneum, and can be absorbed by the body. Pharmacokinetic data, i.e. onthe bioavailability, can be obtained analogously to the method of J.Shaffer et al., J. Pharm. Sciences, 1999, 88, 313–318.

The invention relates to compounds of the formula I according to Claim 1and physiologically acceptable salts and/or solvates thereof astherapeutic active ingredients.

The invention accordingly relates to compounds of the formula Iaccording to Claim 1 and physiologically acceptable salts and/orsolvates thereof as integrin inhibitors.

The invention relates to compounds of the formula I according to Claim 1and physiologically acceptable salts and/or solvates thereof for use incombating diseases.

The compounds of the formula I can be employed as medicament activeingredients in human and veterinary medicine, in particular for theprophylaxis and/or therapy of circulatory diseases, thromboses, cardiacinfarction, arteriosclerosis, apoplexia, angina pectoris, tumourdiseases, such as tumour growth or tumour metastasis, osteolyticdiseases, such as osteoporosis, pathologically angiogenic diseases, suchas, for example, inflammation, ophthalmological diseases, diabeticretinopathy, macular degeneration, myopia, ocular histoplasmosis,rheumatic arthritis, osteoarthritis, rubeotic glaucoma, ulcerativecolitis, Crohn's disease, atherosclerosis, psoriasis, bullate pemphigus,dermatitis, erythema, lung fibrosis, cystic fibrosis, endometriosis,liver cirrhosis, periodontitis, restenosis after angioplasty, multiplesclerosis, viral infections, bacterial infections, fungal infections, inacute renal failure and in wound healing for supporting the healingprocess.

The compounds of the formula l can be employed as antimicrobially activesubstances in operations where biomaterials, implants, catheters orcardiac pacemakers are used. They have an antiseptic action here. Theefficacy of the antimicrobial activity can be demonstrated by the methoddescribed by P. Valentin-Weigund et. al. in Infection and Immunity,1988, 2851–2855.

Since the compounds of the formula I are inhibitors of fibrinogenbinding and are thus ligands of the fibrinogen receptors on bloodplatelets, they can be used in vivo as diagnostic agents for thedetection and localisation of thrombi in vascular systems if they aresubstituted, for example, by a radioactive or UV-detectable radical.

The compounds of the formula I, as inhibitors of fibrinogen binding, canalso be used as effective aids for the study of the metabolism of bloodplatelets in various stages of activation or of intracellular signalmechanisms of the fibrinogen receptor. The detectable unit of a label tobe incorporated, for example isotope labelling by ³H, allows the saidmechanisms to be studied after binding to the receptor.

The following abbreviations are used below:

-   -   Ac acetyl    -   Aza-Gly H₂N—NH—COOH    -   BOC tert-butoxycarbonyl    -   CBZ or Z benzyloxycarbonyl    -   DCCI dicyclohexylcarbodiimide    -   DCM dichloromethane    -   DIPEA diisopropylethylamine    -   DMF dimethylformamide    -   DMSO dimethyl sulfoxide    -   EDCI N-ethyl-N,N′-(dimethylaminopropyl)carbodiimide    -   Et ethyl    -   Fmoc 9-fluorenylmethoxycarbonyl    -   Gly glycine    -   Gua guanidine    -   HATU O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium        hexafluorophosphate    -   HOBt 1-hydroxybenzotriazole    -   Me methyl    -   MBHA 4-methylbenzhydrylamine    -   Mtr 4-methoxy-2,3,6-trimethylphenylsulfonyl    -   NMP N-methylpyrrolidone    -   NMR nuclear magnetic resonance    -   HONSu N-hydroxysuccinimide    -   OBzl benzyl ester    -   OtBu tert-butyl ester    -   Oct octanoyl    -   OMe methyl ester    -   OEt ethyl ester    -   Pbf 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl    -   β-Phe β-phenylalanine    -   POA phenoxyacetyl    -   Pyr pyridine    -   R_(f) value retention factor    -   RP Reversed Phase    -   RT retention time    -   Sal salicyloyl    -   TBTU O-(1H-benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium        tetra-fluoroborate    -   TFA trifluoroacetic acid    -   Thiqu 1,2,3,4-tetrahydroisoquinoline    -   Trt trityl (triphenylmethyl).

The compounds of the formula I have at least one centre of chirality andcan therefore occur in a number of stereoisomeric forms. All these forms(for example D and L forms) and mixtures thereof (for example the DLforms) are included in the formula I.

The compounds according to Claim 1 according to the invention alsoinclude so-called prodrug derivatives, i.e. compounds of the formula Iwhich have been modified with, for example, alkyl or acyl groups, sugarsor oligopeptides and which are rapidly cleaved in the organism to givethe effective compounds according to the invention.

These also include biodegradable polymer derivatives of the compoundsaccording to the invention, as described, for example, in Int. J. Pharm.1995, 115, 61–67.

The compounds according to Claim 1 according to the invention alsoinclude derivatives of the compounds of the formula I whose carboxylgroup has been converted into a pharmaceutically acceptablemetabolically labile ester or an amide thereof.

Furthermore, free amino groups or free hydroxyl groups as substituentsof compounds of the formula I may have been provided with correspondingprotecting groups.

The term solvates of the compounds of the formula I is taken to meanadductions of inert solvent molecules onto the compounds of the formulaI which form owing to their mutual attractive force. Solvates are, forexample, monohydrates or dihydrates or addition compounds with alcohols,such as, for example, with methanol or ethanol.

The invention furthermore relates to a process for the preparation ofcompounds of the formula I according to claim 1 and salts thereof,characterised in that

-   -   a) a compound of the formula II

-   -    in which Z, R¹ and n are as defined above, and W is a        conventional protecting group or a solid phase used in peptide        chemistry,    -    is reacted with a compound of the formula III

-   -    in which Y is as defined above, and Q is a suitable protecting        group or Het, in the presence of a condensing agent, such as,        for example, HATU,    -    and the protecting groups and/or the solid phase are        subsequently removed,    -    and, where appropriate, the resultant product is, if Q as        protecting group is removed, reacted with a suitable guanyl        compound, such as, for example, N,N′-bis-BOC-1-guanylpyrazole,        and, if desired, the remaining protecting groups and/or the        solid phase are removed, or    -   b) a compound of the formula I is liberated from one of its        functional derivatives by treatment with a solvolysing or        hydrogenolysing agent,    -    and/or in that a basic or acidic compound of the formula I is        converted into one of its salts by treatment with an acid or        base.

Throughout the invention, all radicals which occur more than once, suchas, for example, R¹, may be identical or different, i.e. are independentof one another.

In the above formulae, A is alkyl, is linear or branched, and has from 1to 6, preferably 1, 2, 3, 4, 5 or 6 carbon atoms. A is preferablymethyl, furthermore ethyl, isopropyl, n-propyl, n-butyl, isobutyl,sec-butyl or tert-butyl, furthermore also n-pentyl, 1-, 2- or3-methylbutyl, 1,1-, 1,2- or 2,2-dimethylpropyl, 1-ethylpropyl, hexyl,1-, 2-, 3- or4-methylpentyl, 1,1-, 1,2-, 1,3-, 2,2-, 2,3- or3,3-dimethylbutyl, 1- or 2-ethylbutyl, 1-ethyl-1-methylpropyl,1-ethyl-2-methylpropyl, 1,1,2- or 1,2,2-trimethylpropyl. A isparticularly preferably methyl.

The term “protecting group” preferably denotes acetyl, propionyl,butyryl, phenylacetyl, benzoyl, tolyl, POA, methoxycarbonyl,ethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, BOC, 2-iodoethoxycarbonylCBZ (“carbobenzoxy”), 4-methoxybenzyloxycarbonyl, Fmoc, Mtr or benzyl,particularly preferably Fmoc.

Arylalkyl is preferably benzyl, phenylethyl, phenylpropyl ornaphthylmethyl, particularly preferably benzyl.

Hal is preferably F, Cl or Br.

Het is a monocyclic or bicyclic aromatic or saturated radical having upto three heteroatoms, preferably a saturated, partially or completelyunsaturated monocyclic or bicyclic heterocyclic radical having from 5 to10 ring members, where 1 or 2 N and/or 1 or 2 S or O atoms may bepresent and the heterocyclic radical may be monosubstituted ordisubstituted by CN, Hal, OH, OA, CF₃, A, NO₂ or OCF₃.

Het is preferably substituted or unsubstituted 2- or 3-furyl, 2- or3-thienyl, 1-, 2- or 3-pyrrolyl, 1-, 2-, 4- or 5-imidazolyl, 3-, 4- or5-pyrazolyl, 2-, 4- or 5-oxazolyl, 3-, 4- or 5-isoxazolyl, 2-, 4- or5-thiazolyl, 3-, 4- or 5 -isothiazolyl, 2-, 3- or 4-pyridyl, 2-, 4-, 5-or 6-pyrimidinyl, furthermore preferably 1,2,3-triazol-1-, -4- or -5-yl,1,2,4-triazol-1-, -4- or -5-yl, 1- or 5-tetrazolyl, 1,2,3-oxadiazol-4-or -5-yl, 1,2,4-oxadiazol-3- or -5-yl, 1,3,4-thiadiazol-2- or -5-yl,1,2,4-thiadiazol-3- or -5-yl, 1,2,3-thiadiazol-4- or -5-yl, 2-, 3-, 4-,5- or 6-2H-thiopyranyl, 2-, 3- or 4-4H-thiopyranyl, 3- or 4-pyridazinyl,pyrazinyl, 2-, 3-, 4-, 5-, 6- or 7-benzofuryl, 2-, 3-, 4-, 5-, 6- or7-benzothienyl, 1-, 2-, 3-, 4-, 5-, 6- or 7-1H-indolyl, 1-, 2-, 4- or5-benzimidazolyl, 1-, 3-, 4-, 5-, 6-, or 7-benzopyrazolyl, 2-, 4-, 5-,6- or 7-benzoxazolyl, 3-, 4-, 5-, 6- or 7-benzisoxazolyl, 2-, 4-, 5-, 6-or 7-benzothiazolyl, 2-, 4-, 5-, 6-, or 7-benzisothiazolyl, 4-, 5-, 6-or 7-benz-2,1,3-oxadiazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, or8-quinolinyl, 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolinyl, 1-, 2-, 3-, 4-or 9-carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8- or 9-acridinyl, 3-, 4-,5-, 6-, 7- or 8-cinnolinyl, 2-, 4-, 5-, 6-, 7- or 8-quinazolinyl. Theheterocyclic radicals may also be partially or fully hydrogenated. Hetmay thus also be 2,3-dihydro-2-, -3-, -4- or -5-furyl, 2,5-dihydro-2-,-3-, -4- or -5-furyl, tetrahydro-2- or -3-furyl, 1,3-dioxolan-4-yl,tetrahydro-2- or -3-thienyl, 2,3-dihydro-1-, -2-, -3-, -4- or-5-pyrrolyl, 2,5-dihydro-1-, -2-, -3-, 4- or -5-pyrrolyl, 1-, 2- or3-pyrrolidinyl, tetrahydro-1-, -2- or -3-pyrollyl, tetrahydro-1-, -2- or-4-imidazolyl, 2,3-dihydro-1, -2-, -3-, 4-, -5-, -6- or -7-1H-indolyl,2,3-dihydro-1-, -2-, -3-, -4- or -5-pyrazolyl, tetrahydro-1-, -3- or-4-pyrazolyl, 1,4-dihydro-1-, -2-, -3- or -4-pyridyl,1,2,3,4-tetrahydro-1-, -2-, -3-, 4-, -5- or -6-pyridyl,1,2,3,6-tetrahydro-1-, -2-, -3-, 4-, -5- or -6-pyridyl, 1-, 2-, 3- or4-piperidinyl, 1-, 2-, 3- or 4-azepanyl, 2-, 3- or 4-morpholinyl,tetrahydro -2-, -3- or -4-pyranyl, 1,4-dioxanyl, 1,3-dioxan-2-, -4- or-5-yl, hexahydro-1-, -3- or -4-pyridazinyl, hexahydro-1-, -2-, -4- or-5-pyrimidinyl, 1-, 2- or 3-piperazinyl, 1,2,3,4-tetrahydro-1-, -2-,-3-, -4-, -5-, -6-, -7- or -8-quinolinyl, 1,2,3,4-tetrahydro-1-, -2-,-3-,-4-, -5-,-6-, -7- or -8-isoquinolinyl. Het is particularlypreferably methylpyridyl, in particular 4-methylpyridin-2-yl,pyridin-2-yl, pyrimidin-2-yl, imidazol-2-yl, benzimidazol-2-yl andhydrogenated derivatives thereof.

OA is preferably methoxy, ethoxy, propoxy or butoxy, furthermore alsopentyloxy or hexyloxy.

R¹ and R⁵, independently of one another, are preferably H, A, CN, NO₂,Hal or —COA, where A is as defined above; in particular, R¹ and R⁵ areH.

R² is preferably H or A, where A is as defined above; in particular H.

R³ and R⁴, independently of one another, are preferably H or —COA, inparticular H.

X is preferably H, —C(═NH)—NH₂, —C(═N-methyl)-NH₂, 4-methylpyridin-2-yl,pyridin-2-yl, pyrimidin-2-yl, imidazol-2-yl, benzimidazol-2-yl andhydrogenated derivatives thereof.

Y is —(CH₂)_(m)- or

in particular —(CH₂)₄— or

n and p, independently of one another, are preferably 1 or 2, inparticular 1.

m is preferably 0, 2 or 4, in particular 0 or 4.

Preference is given to the compounds of the formulae IA and IB:

in which X, Y, Z and R² are as defined above. R² in the formulae IA andIB is, in particular, H

Accordingly, the invention relates, in particular, to the compounds ofthe formula I in which at least one of the said radicals has one of thepreferred meanings indicated above. Some preferred groups of compoundscan be expressed by the following sub-formulae I1 to I36:

The compounds of the formula I and also the starting materials for theirpreparation are, in addition, prepared by methods known per se, asdescribed in the literature (for example in the standard works, such asHouben-Weyl, Methoden der organischen Chemie [Methods of OrganicChemistry], Georg-Thieme-Verlag, Stuttgart), to be precise underreaction conditions which are known and suitable for the said reactions.Use can also be made here of variants which are known per se, but arenot mentioned here in greater detail.

If desired, the starting materials can also be formed in situ, so thatthey are not isolated from the reaction mixture, but instead areimmediately converted further into the compounds of the formula I.

Compounds of the formula I can preferably be obtained under theconditions of a peptide synthesis. Use is advantageously made here ofconventional methods of peptide synthesis, as described, for example, inHouben-Weyl, 1.c., Volume 15/II, pages 1 to 806 (1974).

The direct precursors of the compounds of the formula I can also bebuilt up on a solid phase, for example a swellable polystyrene resin, asdescribed, for example, by Merrifield (Angew. Chem. 97, 801–812, 1985).Solid phases which can be used are in principle all supports as areknown, for example, from solid-phase peptide chemistry or nucleic acidsynthesis. Suitable polymeric support materials are polymeric solidphases, preferably having hydrophilic properties, for examplecrosslinked polysugars, such as cellulose, sepharose or Sephadex^(R),acrylamides, polyethylene glycol-based polymers or tentaclepolymers^(R).

The solid phase employed is preferably trityl chloride-polystyreneresin, 4-methoxytrityl chloride resin, Merrifield resin or Wang resin.

Thus, compounds of the formula I can be obtained by reacting a compoundof the formula II with a compound of the formula III and subsequentlyremoving the protecting groups or the solid phase.

The compounds of the formula I can likewise, be obtained by reacting acompound of the formula IV with a compound of the formula V andsubsequently removing the protecting groups.

The coupling reaction preferably succeeds in the presence of adehydrating agent, for example a carbodiimide, such as DCCI or EDCI,furthermore, for example, propanephosphonic anhydride (cf: Angew. Chem.1980, 92, 129), diphenylphosphoryl azide or2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline, in an inert solvent, forexample a halogenated hydrocarbon, such as dichloromethane, an ether,such as tetrahydrofuran or dioxane, an amide, such as DMF ordimethylacetamide, a nitrile, such as acetonitrile, in dimethylsulfoxide or in the presence of this solvent, at temperatures betweenabout −10 and 40°, preferably between 0 and 30°. In order to promoteintramolecular cyclisation ahead of intermolecular peptide binding, itis advantageous to work in dilute solutions. The reaction time,depending on the conditions used, is between a few minutes and 14 days.

Instead of compounds of the formulae II and/or IV, it is also possibleto employ derivatives of compounds of the formulae II and/or IV,preferably a pre-activated carboxylic acid, or a carboxylic acid halide,a symmetrical or mixed anhydride or an active ester. Radicals of thistype for activation of the carboxyl group in typical acylation reactionshave been described in the literature (for example in the standardworks, such as Houben-Weyl, Methoden der organischen Chemie [Methods ofOrganic Chemistry], Georg-Thieme-Verlag, Stuttgart). Activated estersare advantageously formed in situ, for example by addition of HOBt orN-hydroxysuccinimide.

The reaction is generally carried out in an inert solvent; if acarboxylic acid halide is used, it is carried out in the presence of anacid-binding agent, preferably an organic base, such as triethylamine,dimethylaniline, pyridine or quinoline.

The addition of an alkali or alkaline-earth metal hydroxide, carbonateor bicarbonate or another salt of a weak acid of the alkali oralkaline-earth metals, preferably of potassium, sodium, calcium orcaesium, may also be favourable.

The compounds of the formula I can furthermore be obtained by liberatingthem from their functional derivatives by solvolysis, in particularhydrolysis, or by hydrogenolysis.

Preferred starting materials for the solvolysis or hydrogenolysis arethose which otherwise conform to the formula I, but in which one or morefree amino and/or hydroxyl groups have been replaced by correspondingprotected amino and/or hydroxyl groups, in particular those in which anH—N group has been replaced by an SG¹—N group, in which SG¹ is anamino-protecting group, and/or those in which the H atom of a hydroxylgroup has been replaced by a hydroxyl-protecting group, for examplethose which conform to the formula I, but in which a —COOH group hasbeen replaced by a —COOSG² group, in which SG² is a hydroxyl-protectinggroup.

It is also possible for a plurality of—identical or different—protectedamino and/or hydroxyl groups to be present in the molecule of thestarting material. If the protecting groups present differ from oneanother, they can in many cases be removed selectively (cf. in thisrespect: T. W. Greene, P. G. M. Wuts, Protective Groups in OrganicChemistry, 2nd Edn., Wiley, New York 1991 or P. J. Kocienski, ProtectingGroups, 1st Edn., Georg Thieme Verlag, Stuttgart-New York, 1994), H.Kunz, H. Waldmann in Comprehensive Organic Synthesis, Vol. 6 (Eds. B. M.Trost, I. Fleming, E. Winterfeldt), Pergamon, Oxford, 1991, pp.631–701).

The term “amino protecting group” is generally known and relates togroups which are suitable for protecting (blocking) an amino groupagainst chemical reactions. Typical of such groups are, in particular,unsubstituted or substituted acyl, aryl, aralkoxymethyl or aralkylgroups. Since the amino protecting groups are removed after the desiredreaction (or synthesis sequence), their type and size is furthermore notcrucial; however, preference is given to those having 1–20 carbon atoms.The term “acyl group” is to be understood in the broadest sense inconnection with the present process. It includes acyl groups derivedaliphatic, araliphatic, alicyclic, aromatic and heterocyclic carboxylicacids or sulfonic acids, as well as, in particular, alkoxycarbonyl,alkenyloxycarbonyl, aryloxycarbonyl and especially aralkoxycarbonylgroups. Examples of such acyl groups are alkanoyl, such as acetyl,propionyl and butyryl; aralkanoyl, such as phenylacetyl; aroyl, such asbenzoyl and tolyl; aryloxyalkanoyl, such as phenoxyacetyl;alkoxycarbonyl, such as methoxycarbonyl, ethoxycarbonyl,2,2,2-trichloroethoxycarbonyl, Boc and 2-iodoethoxycarbonyl;alkenyloxycarbonyl, such as allyloxycarbonyl (Aloc), aralkoxycarbonyl,such as CBZ (synonymous with Z), 4-methoxybenzyloxycarbonyl (MOZ),4-nitrobenzyloxycarbonyl and 9-fluorenylmethoxycarbonyl (Fmoc);2-(phenylsulfonyl)ethoxycarbonyl; trimethylsilylethoxycarbonyl (Teoc),and arylsulfonyl, such as 4-methoxy-2,3,6-trimethylphenylsulfonyl (Mtr).Preferred amino protecting groups are Boc, Fmoc and Aloc, furthermore Z,benzyl and acetyl.

The term “hydroxyl protecting group” is likewise generally known andrelates to groups which are suitable for protecting a hydroxyl groupagainst chemical reactions. Typical of such groups are theabove-mentioned unsubstituted or substituted aryl, aralkyl, aroyl oracyl groups, furthermore also alkyl groups, alkyl-, aryl- andaralkylsilyl groups, and O,O— and O,S-acetals. The nature and size ofthe hydroxyl protecting groups is not crucial since they are removedagain after the desired chemical reaction or synthesis sequence;preference is given to groups having 1–20 carbon atoms, in particular1–10 carbon atoms. Examples of hydroxyl protecting groups are, interalia, aralkyl groups, such as benzyl, 4-methoxybenzyl and2,4-dimethoxybenzyl, aroyl groups, such as benzoyl and p-nitrobenzoyl,acyl groups, such as acetyl and pivaloyl, p-toluenesulfonyl, alkylgroups, such as methyl and tert-butyl, but also allyl, alkylsilylgroups, such as trimethylsilyl (TMS), triisopropylsilyl (TIPS),tert-butyldimethylsilyl (TBS) and triethylsilyl, trimethylsilylethyl,aralkylsilyl groups, such as tert-butyldiphenylsilyl (TBDPS), cyclicacetals, such as isopropylidene acetal, cyclopentylidene acetal,cyclohexylidene acetal, benzylidene acetal, p-methoxybenzylidene acetaland o,p-dimethoxybenzylidene acetal, acyclic acetals, such astetrahydropyranyl (Thp), methoxymethyl (MOM), methoxyethoxymethyl (MEM),benzyloxymethyl (BOM) and methylthiomethyl (MTM). Particularly preferredhydroxyl protecting groups are benzyl, acetyl, tert-butyl and TBS.

The liberation of the compounds of the formula I from their functionalderivatives is known from the literature for the protecting group usedin each case (for example T. W. Greene, P. G. M. Wuts, Protective Groupsin Organic Chemistry, 2nd Edn., Wiley, New York 1991 or P. J. Kocienski,Protecting Groups, 1st Edn., Georg Thieme Verlag, Stuttgart-New York,1994). Use may also be made here of variants which are known per se, butare not mentioned here in greater detail.

A base of the formula I can be converted into the associatedacid-addition salt using an acid, for example by reaction of equivalentamounts of the base and the acid in an inert solvent, such as ethanol,followed by evaporation. Suitable acids for this reaction are, inparticular, those which give physiologically acceptable salts. Thus, itis possible to use inorganic acids, for example sulfuric acid, sulfurousacid, dithionic acid, nitric acid, hydrohalic acids, such ashydrochloric acid or hydrobromic acid, phosphoric acids, such as, forexample, orthophosphoric acid, sulfamic-acid, furthermore organic acids,in particular aliphatic, alicyclic, araliphatic, aromatic orheterocyclic monobasic or polybasic carboxylic, sulfonic or sulfuricacids, for example formic acid, acetic acid, propionic acid, hexanoicacid, octanoic acid, decanoic acid, hexadecanoic acid, octadecanoicacid, pivalic acid, diethylacetic acid, malonic acid, succinic acid,pimelic acid, fumaric acid, maleic acid, lactic acid, tartaric acid,malic acid, citric acid, gluconic acid, ascorbic acid, nicotinic acid,isonicotinic acid, methane- or ethanesulfonic acid, benzenesulfonicacid, trimethoxybenzoic acid, adamantanecarboxylic acid,p-toluenesulfonic acid, glycolic acid, embonic acid,chlorophenoxy-acetic acid, aspartic acid, glutamic acid, proline,glyoxylic acid, palmitic acid, para-chlorophenoxyisobutyric acid,cyclohexanecarboxylic acid, glucose 1-phosphate, naphthalenemono- and-disulfonic acids or laurylsulfuric acid. Salts with physiologicallyunacceptable acids, for example picrates, can be used to isolate and/orpurify the compounds of the formula I. On the other hand, compounds ofthe formula I can be converted into the corresponding metal salts, inparticular alkali metal salts or alkaline earth metal salts, or into thecorresponding ammonium salts, using bases (for example sodium hydroxide,potassium hydroxide, sodium carbonate or potassium carbonate). Suitablesalts are furthermore substituted ammonium salts, for example thedimethyl-, diethyl- or diisopropyl-ammonium salts, monoethanol-,diethanol- or diisopropylammonium salts, cyclohexyl- ordicyclohexylammonium salts, dibenzylethylenediammonium salts,furthermore, for example, salts with arginine or lysine

The invention furthermore relates to the use of the compounds of theformula I and/or physiologically acceptable salts thereof for thepreparation of a medicament.

The invention furthermore relates to pharmaceutical preparationscomprising at least one compound of the formula I and/or one of itsphysiologically acceptable salts or solvates thereof which are prepared,in particular, by non-chemical methods. In this case, the compounds ofthe formula I can be brought into a suitable dosage form here togetherwith at least one solid, liquid and/or semi-liquid excipient or adjuvantand, if desired, in combination with one or more further activeingredients.

These preparations can be used as medicaments in human or veterinarymedicine. Suitable excipients are organic or inorganic substances whichare suitable for enteral (for example oral), parenteral or topicaladministration and do not react with the novel compounds, for examplewater, vegetable oils, benzyl alcohols, alkylene glycols, polyethyleneglycols, glycerol triacetate, gelatine, carbohydrates, such as lactoseor starch, magnesium stearate, talc or vaseline. Suitable for oraladministration are, in particular, tablets, pills, coated tablets,capsules, powders, granules, syrups, juices or drops, suitable forrectal administration are suppositories, suitable for parenteraladministration are solutions, preferably oily or aqueous solutions,furthermore suspensions, emulsions or implants, and suitable for topicalapplication are ointments, creams or powders. The novel compounds canalso be lyophilised and the resultant lyophilisates used, for example,for the preparation of injection preparations. The preparationsindicated may be sterilised and/or comprise assistants, such aslubricants, preservatives, stabilisers and/or wetting agents,emulsifiers, salts for modifying the osmotic pressure, buffersubstances, dyes, flavours and/or a plurality of further activeingredients, for example one or more vitamins. For administration as aninhalation spray, it is possible to use sprays in which the activeingredient is either dissolved or suspended in a propellant gas orpropellant gas mixture (for example CO₂ or chlorofluorocarbons). Theactive ingredient is advantageously used here in micronised form, inwhich case one or more additional physiologically acceptable solventsmay be present, for example ethanol. Inhalation solutions can beadministered with the aid of conventional inhalers.

The compounds of the formula I and physiologically acceptable saltsthereof can be used as integrin inhibitors in the combating of diseases,in particular thromboses, cardiac infarction, coronary heart diseases,arteriosclerosis, tumours, osteoporosis, inflammation and infections.

The compounds of the formula I and physiologically acceptable saltsthereof can also be used in the case of pathological processesmaintained or propagated by angiogenesis, in particular in the case oftumours or rheumatoid arthritis.

The substances according to the invention are generally administeredanalogously to other known, commercially available peptides, but inparticular analogously to the compounds described in U.S. Pat. No.4,472,305, preferably in doses of from about 0.05 to 500 mg, inparticular from 0.5 to 100 mg, per dosage unit. The daily dose ispreferably from about 0.01 to 2 mg/kg of body weight. However, thespecific dose for each patient depends on a wide variety of factors, forexample on the efficacy of the specific compound employed, on the age,body weight, general state of health, sex, on the diet, on the time andmethod of administration, on the rate of excretion, medicamentcombination and severity of the particular disease to which the therapyapplies. Parenteral administration is preferred.

Furthermore, the compounds of the formula I can be used as integrinligands for the production of columns for affinity chromatography forthe purification of integrins.

In this method, the ligand, i.e. a compound of the formula I, iscovalently coupled to a polymeric support via an anchor function, forexample the carboxyl group of Asp.

The materials for affinity chromatography for integrin purification areprepared under conditions as are usual and known per se for thecondensation of amino acids.

The compounds of the formula I have one or more centres of chirality andcan therefore exist in racemic or optically active form. Racematesobtained can be resolved into the enantiomers mechanically or chemicallyby methods known per se. Diastereomers are preferably formed from theracemic mixture by reaction with an optically active resolving agent.Examples of suitable resolving agents are optically active acids, suchas the D and L forms of tartaric acid, diacetyltartaric acid,dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, and thevarious optically active camphorsulfonic acids, such asβ-camphorsulfonic acid. Resolution of the enantiomers with the aid of acolumn filled with an optically active resolving agent (for exampledinitrobenzoylphenylglycine) is also advantageous; an example of asuitable eluent is a mixture of hexane/isopropanol/acetonitrile, forexample in the volume ratio 82:15:3.

It is of course also possible to obtain optically active compounds ofthe formula I by the methods described above by using starting materialswhich are already optically active.

Above and below, all temperatures are given in ° C. In the followingexamples, “conventional work-up” means that, if necessary, water isadded, if necessary, depending on the constitution of the end product,the pH is adjusted to a value between 2 and 10, the mixture is extractedwith ethyl acetate or dichloromethane, the phases are separated, theorganic phase is dried over sodium sulfate and evaporated, and theproduct is purified by chromatography on silica gel and/or bycrystallisation.

RT=retention time (minutes) in HPLC in the following systems:

-   -   Columns from Omnicrom YMC:    -   1. 4.6×250 mm, 5 μm, C₁₈ (analysis);    -   2. 30×250 mm, 7 μm, C₁₈ (preparation).        The eluents used are gradients comprising acetonitrile (B) with        0.1% of TFA and water (A) with 0.1% of TFA (data in each case in        per cent by volume of acetonitrile). The retention time RT was        determined at a flow rate of 1 ml/min.        Detection at 220 nm.

The diastereomers are preferably separated under the stated conditions.

-   -   Mass spectrometry (MS): ESI (electrospray ionisation) (M+H)⁺.        -   FAB (fast atom bombardment) (M+H)⁺.            1. Material and General Working Procedures

Solvents for the synthesis were either obtained in “technical grade” anddistilled before use or purchased from Fluka (Seelze) or Merck(Darmstadt) in purity grades “absolute” or “for synthesis”. NMP(distilled) was obtained free of charge from BASF (Ludwigshafen).Solvents for column chromatography were obtained in “technical grade”and either distilled before use or employed without distillation(hexane). The HPLC solvents acetonitrile (solvent B) and TFA werepurchased in purity grade “gradient grade” from Merck (Darmstadt), water(solvent A) was deionised and treated with a Milli-Q system fromMillipore (Molsheim, France).

Fmoc-protected amino acids were purchased from Novabiochem, AdvancedChemTech, MultiSynTech or PepTech Corporation (Cambridge, USA).

For manual solid-phase synthesis, use was made of PE syringes fromBecton-Dickinson (Fraga, Spain) or Braun (Melsungen) with PE frits fromRoland Vetter Laborbedarf (Ammerbuch). In order to mix the resinsuspension, the syringes were rotated at about 30 rpm. The resin wascharged in glass shaking vessels.

Air- or moisture-sensitive reactions were carried out in dry glassvessels and under an argon atmosphere (99.996%). Hygroscopic solventsand/or solvents which had been rendered absolute were transferred intosyringes under argon.

For the HPLC purification, the compounds were dissolved in DMSO,acetonitrile or methanol (“gradient grade”) and filtered through an RC15 or RC 25 (RC membrane, 0.45 μm) syringe filter from Sartorius(Göttingen). Analytical, semi-preparative and preparative separationswere carried out in two HPLC systems from Amersham Pharmacia Biotech(analytical: Äkta Basic 10F with A-900 autosampler; preparative: ÄktaBasic 100F with P-900 pump system and UV-900 detector) and two systemsfrom Beckman (Gold system with 125 solvent module and 166 detectormodule; 110B pump system, 420 control unit and Knauer Uvicord detector).The following columns were used for analytical separations: ODS-A C₁₈(250 mm×4.6 mm, 5 μm, flow rate: 1 ml/min) from Omnicrom YMC; forsemi-preparative separations: ODS-A C₁₈ (250 mm×20 mm, 5 μm or 10 μm,flow rate: 8 ml/min) from Omnicrom YMC; for preparative separations:ODS-A C₁₈ (250 mm×30 mm, 10 μm, flow rate: 25 ml/min) from Omnicrom YMCand Nucleosil C₁₈ (250 mm×20 mm, 7 μm, flow rate: 25 ml/min) fromMacherey-Nagel. The compounds were eluted with linear gradients (30 min)of acetonitrile (solvent B) in water (solvent A) and 0.1% (v/v) of TFA.For analytical purity determination of the compounds aftersemi-preparative or preparative HPLC purification, the peak integral ofthe analytical HPLC chromatogram was evaluated at a detector wavelengthof 220 nm.

The column chromatography was carried out using silica gel 60 (230–400mesh ASTM, particle size 0.040–0.063 mm) from Merck (Darmstadt), flashchromatography was carried out at a pressure of 1–1.2 bar aboveatmospheric.

Thin-layer chromatography (TLC) and the determination of the R_(f)values were carried out using aluminium TLC plates coated with silicagel 60 F₂₅₄ from Merck (Darmstadt). For detection, the TLC plates wereviewed under UV light (λ=254 nm).

Melting points were determined by the Dr Tottoli method in a Büchi 510melting point apparatus and are uncorrected.

All ¹H-NMR and ¹³C-NMR spectra were recorded on a Bruker AC250 or DMX500spectrometer at 300 K, the spectral data were processed on Bruker Aspekt1000 (AC 250) or on Silicon Graphics Indy, O2 and Octane workstationswith XWINNMR software. Chemical shifts (δ) are given in parts permillion (ppm) relative to tetramethylsilane, and coupling constants aregiven in hertz (Hz). The internal standard used was tetramethylsilane orthe solvent peak: DMSO-d₆: 2.49 ppm (¹H-NMR) and 39.5 ppm (¹³C-NMR);CDCl₃: 7.24 ppm (¹H-NMR) and 77.0 ppm (¹³C-NMR). ¹³C-NMR spectra wererecorded with ¹H broad-band decoupling. The signal assignment was inmost cases carried out with the aid of HMQC and COSY experiments.

Mass spectra were recorded by the electron impact (El) and chemicalionisation (Cl) techniques on a Finnigan MAT 8200 instrument.Electrospray ionisation (ESI) mass spectra were recorded on a FinniganLCQ mass spectrometer in combination with a Hewlett Packard 1100 HPLCsystem with an ODS-A C₁₈ (125 mm×2 mm, 3 μm, flow rate: 0.2 ml/min)column from Omnicrom YMC. The compounds were eluted with a lineargradient (15 min) of acetonitrile (solvent B) in water (solvent A) and0.1% (v/v) of formic acid. Mass spectra are given in the form “X (Y)[M+Z]⁺”, where “X” is the detected mass, “Y” is the observed intensityof the mass peak, “M” is the molecule investigated, and “Z” is theadducted cation.

High-resolution mass spectra (HRMS) were recorded by Koka JajasimhuluPh.D. (University of Cincinnati, USA) using the electrosprayionisation-time of flight (ESI-TOF) technique.

Lyophilisation was carried out using the Alpha 2–4 instrument fromChrist (Osterode).

AAV1: Loading of TCP Resin

The corresponding Fmoc-protected amino acid (1.56 mmol, 1.5 eq) andDIPEA (177 μl, 1.03 mmol) are added to pre-swollen TCP resin (1.16 g,maximum loading: 0.9 mmol/g) in dry CH₂Cl₂ (6 ml, 10 min). After 5minutes, further DIPEA (91 μl, 0.52 mmol) is added, and the resin isshaken. After 2 hours, methanol (1.16 ml) is added in order to cap theunreacted trityl groups, and the resin is shaken for a further 15minutes. The resin is then washed with dry CH₂Cl₂ (5×20 ml, 3 minuteseach time), NMP (5×20 ml, 3 minutes each time) and again with dry CH₂Cl₂(5×20 ml, 3 minutes each time) and finally with a mixture-ofmethanol/CH₂Cl₂ (1:1, 20 ml) and methanol (20 ml). The resin is dried ina high vacuum, and the loading can be determined using the followingequation:

$l = \frac{\left( {m_{2} - m_{1}} \right) \times 1000}{\left( {{MW} - 36.45} \right) \times m_{2}}$

-   -   l loading of the resin with unit [mmol/g]    -   m₁ weight of the resin before coupling [g]    -   m₂ weight of the dried resin after coupling [g]    -   MW molecular weight of the Fmoc-protected amino acid/carboxylic        acid unit [g/mol]

The error arising through the difference masses of Cl and MeO can beneglected.

AAV 2: Removal of the Fmoc Protecting Group

The resin (100 mg) is pre-swollen in NMP (5 ml, 10 min). The Fmocprotecting group is removed by treatment with a freshly prepared 20%piperidine solution (v/v) in NMP (5 ml) for 15 minutes. The resin isthen washed with NMP (5×5 ml, 3 minutes each time), and a 20% piperidinesolution (v/v) in NMP (5 ml, 15 minutes) is again added. Finally, theresin is washed with NMP (5×5 ml, 3 minutes each time).

AAV 3: Coupling of 5-(9H-fluoren-9-ylmethoxy)-3H-1,3,4-oxadiazol-2-one(142) to Resin-Bound, Free Amines by the Gibson Method

In order to deprotect the resin-bound amine, 20% piperidine (v/v) in NMP(2×5 ml, 15 minutes each time) is added to the TCP resin (100 mg, 0.354mmol/g, 0.035 mmol). The resin is then washed with NMP (5×5 ml, 3minutes each time) and dry CH₂Cl₂ (5×5 ml, 3 minutes each time) and thenswollen in dry CH₂Cl₂ (5 ml) for half an hour. A solution of5-(9H-fluoren-9-ylmethoxy)-3H-1,3,4-oxadiazol-2-one (142) (30.5 mg,0.108 mmol, 3.1 eq) in dry CH₂Cl₂ (1 ml) is then added to the resin, andthe mixture is shaken for 90 minutes. The reaction is terminated bywashing with CH₂Cl₂ (5×5 ml, 3 minutes each time) and NMP (5×5 ml, 3minutes each time).

AAV 4: Coupling with HATU/HOAt

The resin-bound, free amine or hydrazine (0.389 mmol) is washed with NMP(5×5 ml, 3-minutes each time). A solution of the suitable Fmoc-protectedamino acid or of a carboxylic acid unit (0.779 mmol, 2 eq), HATU (296mg, 0.779 mmol, 2 eq) and HOAt (106 mg, 0.779 mmol, 2 eq) in NMP (5 ml)is then added to the resin. Finally, sym-collidine (1027 μl, 7.79 mmol,20 eq) is added, and the resin is shaken overnight. The resin is thenwashed with NMP (5×5 ml, 3 minutes each time), and the coupling step isrepeated with the same reagents, amounts and reaction time. The resin issubsequently washed with NMP (5×5 ml, 3 minutes each time).

AAV 5: Removal from the TCP Resin

The compound is removed from the TCP resin in accordance with thefollowing flow chart:

Step Reagents Operation Number Time [min] 1 CH₂Cl₂ washing 3 10 2TFA/TIPS/H₂O removal/deprotection 3 30 (18:1:1) 3 CH₂Cl₂ washing 3 3

For 100 mg of resin, 2 ml of removal solution were usually used. Thecombined filtrates from steps 2 and 3 were evaporated.

2. EXAMPLES Example 1a)

N-[(9H-Fluoren-9-ylmethoxy)carbonyl]hydrazine (141)

Boc-hydrazine (10.0 g, 75.6 mmol) and DIPEA (12.95 ml, 75.6 mmol) weredissolved in dry CH₂Cl₂ (200 ml) and cooled to 0° C. FmocCl (19.6 g,75.8 mmol), dissolved in dry CH₂Cl₂ (100 ml), was then added over thecourse of 30 minutes, and the mixture was stirred overnight at roomtemperature. The organic phase was extracted with water (200 ml) andevaporated to a volume of about 100 ml. Trifluoroacetic acid (100 ml)was then carefully added at 0° C., and the mixture was stirred for 1.5hours. The product was precipitated by careful addition of saturatedNa₂CO₃ solution (300 ml) and dried, giving a colourless solid (18.02 g,70.8 mmol, 94%).

m.p. 150–153° C.; ¹H-NMR (250 MHz, DMSO-d₆, 300 K) δ=10.10 (bs, 1H, NH),9.60 (bs, 1H, NH), 7.89 (d, J=7.6 Hz, 2H, arom), 7.70 (d, J=7.3 Hz, 2H,arom), 7.30–7.45 (m, 4H, arom), 4.48 (d, J=6.6 Hz, 2H, CO—CH₂), 4.27 (t,J=6.7 Hz, 1H, CO—CH₂—CH); ¹³C-NMR (62.9 MHz, DMSO-d₆, 300 K) δ=156.26,143.59, 140.98, 127.96, 127.34, 125.33, 120.39, 67.00, 46.60; HRMS(ESI-TOF) for C₁₅H₁₅N₂O₂ [M+H]⁺: 255.1134 (calc. 255.1119); analyticalHPLC (5–90% in 30 min) t_(R)=16.47 min.

Example 1b)

5-(9H-Fluoren-9-ylmethoxy)-3H-1,3,4-oxadiazol-2-one (142)

A suspension of N-[(9H-fluoren-9-ylmethoxy)carbonyl]hydrazine (141)(1.49 g, 5.78 mmol), CH₂Cl₂ (60 ml) and saturated NaHCO₃ solution (60ml) was stirred vigorously at 0° C. for 5 minutes, and the solution wasthen left for 5 minutes without stirring. Phosgene (1.89 M in toluene,7.95 ml, 15.0 mmol) was then carefully added to the lower, organic phaseusing a syringe, and stirring of the reaction mixture was begun againimmediately after the addition. After 10 minutes, water (20 ml) andCH₂Cl₂ (20 ml) were added to the reaction mixture. The phases were thenseparated rapidly, the aqueous phase was extracted with CH₂Cl₂ (50 ml),and the combined organic phases were dried over Na₂SO₄. Removal of thesolvent under reduced pressure and drying gave a colourless solid (1.35g, 4.82 mmol, 83%).

m.p. 125° C.; ¹H-NMR (250 MHz, CDCl₃, 300 K) δ=8.72 (bs, 1H, NH), 7.77(d, J=7.5 Hz, 2H, arom), 7.59 (d, J=7.4 Hz, 2H, arom), 7.28–7.45 (m, 4H,arom), 4.49 (d, J=7.8 Hz, 2H, CH₂—CH), 4.32–4.41 (m, 1H, CH₂—CH).

Example 2a)

N-Phenylethylformamide (146)

A mixture of phenylethylamine (20.0 g, 0.165 mol) and formic acid (49.4ml, 1.309 mol) was slowly heated to 200° C. Excess water and formic acidwere distilled off in the process. The mixture was then kept at 200° C.for 1 hour, and the product was distilled under reduced pressure, givinga colourless oil (22.0 g, 0.147 mol, 89%).

¹H-NMR (250 MHz, DMSO-d₆, 300 K) δ=8.06 (bs, 1H, CHO), 7.16–7.32 (m, 5H,arom), 3.32–3.42 (m, 2H, NH—CH₂), 2.77 (t, J=7.2 Hz, 2H, NH—CH₂—CH₂);analytical HPLC (5–90% in 30 min) t_(R)=15.04 min.

Example 2b)

3,4-Dihydroisoquinoline (147)

Polyphosphoric acid (25 g) and phosphorus pentoxide (5.4 g, 38.0 mmol)were heated to 180° C. over the course of one hour in an oil bath underan argon atmosphere. N-Phenylethylformamide (146) (4.3 g, 28.8 mmol) wasthen added at 160° C., and the mixture was stirred for 1.5 hours atconstant temperature. The mixture was then allowed to cool to roomtemperature, and water (40 ml) was added. The mixture was subsequentlyadjusted to pH 10 by careful addition of saturated aqueous NaOHsolution. The mixture was then extracted with ether (500 ml), and theorganic phase was separated off and dried using NaOH. Evaporation gave abrown oil (2.81 g, 21.4 mmol, 74%).

¹H-NMR (250 MHz, DMSO-d₆, 300 K) δ=8.32 (t, J=2.3 Hz, 1H, N—CH),7.16–7.40 (m, 4H, arom), 3.59–3.66 (m, 2H, N—CH₂), 2.66 (t, J=7.3 Hz,2H, N—CH₂—CH₂); analytical HPLC (5–90% in 30 min) t_(R)=8.29 min.

Example 2c)

2-(1,2,3,4-Tetrahydro-1-isoquinolinyl)acetic acid (148)

3,4-Dihydroisoquinoline (147) (2.2 g, 16.77 mmol) and malonic acid (1.94g, 16.77 mmol) were mixed at room temperature and heated at 120° C. for1 hour in an oil bath. The mixture was then allowed to cool to roomtemperature, and the product was recrystallised from methanol (150 ml).Drying gave a colourless solid (1.52 g, 7.99 mmol, 48%).

m.p. 230° C. decomp; ¹H-NMR (250 MHz, D₂O, 300 K) δ=7.13–7.23 (m, 4H,arom), 4.65 (t, J=6.9 Hz, 1H, CH—NH), 3.44–3.54 (m, 1H, NH—CH₂),3.24–3.34 (m, 1H, NH—CH₂), 2.95–3.03 (m, 2H, NH—CH₂—CH₂), 2.79 (d, J=6.1Hz, 2H, CH₂—COOH); HRMS (ESI-TOF) for C₁₁H₁₄NO₂ [M+H]⁺: 192.1047 (calc.192.1025); analytical HPLC (5–90% in 30 min) t_(R)=10.15 min.

Example 2d)

9H-Fluoren-9-ylmethyl1-carboxymethyl-3,4-dihydro-1H-isoquinoline-2-carboxylate (149)

A suspension of 2-(1,2,3,4-tetrahydro-1-isoquinolinyl)acetic acid (148)(0.9 g, 4.73 mmol), saturated NaHCO₃ solution (15 ml) and dioxane (5 ml)was cooled to 0° C. FmocCl (1.35 g, 5.2 mmol), dissolved in dioxane (5ml), was then added dropwise over the course of 30 minutes, and themixture was stirred overnight. The mixture was then washed by shakingwith ether (30 ml), and the aqueous phase was adjusted to pH 1 usingconc. HCl. The product was then extracted with ethyl acetate (50 ml),and the organic phase was dried over MgSO₄ and evaporated. The crudeproduct was subsequently purified by column chromatography (ethylacetate/hexane/acetic acid, 1:1:1%) and dried, giving a colourless foam(1.54 g, 3.73 mmol, 79%).

m.p. 61–63° C.; TLC R_(f) (ethyl acetate/hexane/acetic acid,1:1:1%)=0.54; ¹H-NMR (250 MHz, DMSO-d₆, 300 K) δ=7.79–7.91 (m, 2H,arom), 7.63–7.66 (m, 2H, arom), 7.05–7.39 (m, 8H, arom), 5.40–5.52 (m,1H, NH—CH), 4.37–4.42 (m, 1H, COO—CH₂—CH), 4.25–4.32 (m, 2H, COO—CH₂),3.62–4.02 (m, 1H, N—CH₂), 3.27–3.38 (m, 1H, N—CH₂), 2.52–2.76 (m, 4H,N—CH—CH₂ and N—CH₂—CH₂); MS (ESI) m/e 179.1 (30), 414.0 (20) [M+H]⁺,436.2 (25) [M+Na]⁺, 492.8 (15), 826.7 (5) [2M+H]⁺, 849.1 (45) [2M+Na]⁺,865.1 (100) [2M+K]⁺; HRMS (ESI-TOF) for C₂₆H₂₄NO₄ [M+H]⁺: 414.1721(calc. 414.1705); analytical HPLC (5–90% in 30 min) t_(R)=27.53 min.

Example 3a)

Ethyl 5-[N-(4-methylpyridin-2-yl)amino]pentanoate

Ethyl 5-bromopentanoate (33.03 g, 25 ml, 158 mmol) and2-amino-4-methylpyridine (32.9 g, 304 mmol) were refluxed overnight at130° C. (oil-bath temperature). After cooling to room temperature,saturated NaHCO₃ solution (100 ml) was added to the reaction mixture,which was then extracted with ether (5×100 ml). The combined organicphases were dried over MgSO₄, and the solvent was removed. The crudeproduct was purified by flash chromatography (ethyl acetate/hexane, 1:1,2 l; 3:2, 1 l; 7:3, 1 l; 4:1, 1 l), and dried, giving a colourless solid(16.7 g, 70.7 mmol, 45%).

m.p. 41–43° C.; TLC R_(f) (ethyl acetate/hexane, 1:1)=0.26.; ¹H-NMR (250MHz, DMSO-d₆, 300 K) δ=7.90 (d, J=5.3 Hz,1H, N—CH—CH), 6.38 (d, J=5.2Hz, 1H, N—C—CH), 6.17 (s, 1H, N—CH—CH), 4.51 (bs, 1H, NH), 4.11 (q,J=7.2 Hz, 2H, O—CH₂—CH₃), 3.25 (q, J=6.3 Hz, 2H, NH—CH₂), 2.33 (t, J=7.0Hz, 2H, CH₂—CO), 2.21 (s, 3H, C—CH₃),1.60–1.80 (m, 4H, NH—CH₂—CH₂—CH₂),1.23 (t, J=7.0 Hz, 2H, O—CH₂—CH₃); analytical HPLC (5–90% in 30 min)t_(R)=13.58 min.

Example 3b)

5-[N-(4-Methylpyridin-2-yl)amino]pentanoic acid

Ethyl 5-[N-(4-methylpyridin-2-yl)amino]pentanoate (16.7 g, 70.7 mmol,obtainable in accordance with Example 3a)) was dissolved in methanol (20ml), 2N aqueous NaOH (71 ml, 141 mmol) was added, and the mixture wasstirred overnight at room temperature. The solvent was then removed, andthe resultant solid was extracted thoroughly with CHCl₃ (500 ml) and anexcess of DIPEA. The filtrate was evaporated and dried, giving acolourless solid (3.92 g, 18.8 mmol, 27%).

m.p. 138–140° C; ¹H-NMR (250 MHz, DMSO-d₆, 300 K) δ=7.79 (d, J=5.2 Hz,1H, NH), 6.26–6.30 (m, 2H, arom. C⁵—H and C⁶—H), 6.22 (s, 1H, arom.C³—H), 3.17 (dt, J=5.8 Hz, 2H, NH—CH₂), 2.21 (t, J=7.0 Hz, 2H,CH₂—CH₂—CO), 2.11 (s, 3H, C^(quat.)—CH₃), 1.41–1.58 (m, 4H,CH₂—CH₂—CH₂—CH₂); ¹³C-NMR (67.5 MHz, DMSO-d₆, 300 K) δ=174.6 (COOH),159.3, 147.3, 146.8, 113.1, 107.9, 40.5, 33.7, 28.8, 22.4, 20.8; HRMS(ESI-TOF) for C₁₁H₁₇N₂O₂ [M+H]⁺: 209.1293 (calc. 209.1290); analyticalHPLC (5–90% in 30 min) t_(R)=9.83 min.

Example 4

TCP resin was loaded with 9H-fluoren-9-ylmethyl1-carboxymethyl-3,4-dihydro-1H-isoquinoline-2-carboxylate (149 fromExample 2d)) (0.48 g, 1.16 mmol) as described in AAV 1 (m₁=0.84 g,m₂=1.0 g, l=0.426 mmol/g). Fmoc deprotection and coupling of the freshlyprepared 5-(9H-fluoren-9-ylmethoxy)-3H-1,3,4-oxadiazol-2-one (142 fromExample 1b)) (385 mg, 1.32 mmol) were carried out as described in AAV 2and 3, coupling of 5-[N-(4-methylpyridin-2-yl)amino]pentanoic acid (177mg, 0.852 mmol) was carried out as described in AAV 4, and removal ofresin was carried out in accordance with AAV 5. After HPLC purification(10–80% in 30 min) and lyophilisation, a colourless powder was obtained(3.0 mg, 0.00542 mmol, 1.3%).

m.p. 103–109° C.; ¹H-NMR (500 MHz, DMSO-d₆, 300 K) δ=8,81 (bs, 1H,NH—NH), 8.44 (bs, 1H, NH—NH), 8.25 (d, J=6.5 Hz, 1H, N—CH—CH), 7.74–7.77(m, 4H, arom), 7.34 (s, 1H, N—C—CH), 7.21 (d, J=6.4 Hz, 1H, N—CH—CH),6.06–6.07 (m, 1H, N—CH), 4.46–4.52 (m, 1H, CH—CH₂), 3.82–3.93 (m, 3H,NH—CH₂ and CH—CH₂), 3.47–3.57 (m, 2H, NCH₂CH₂), 3.30–3.40 (m, 2H,NCH₂CH₂), 2.94 (s, 3H, C—CH₃), 2.79–2.84 (m, 2H, CH₂—CO), 2.23–2.35 (m,4H, NH—CH₂—CH₂—CH₂); MS (ESI) m/e 191.2 (8), 249.1 (100), 440.1 (30)[M+H]⁺, 462.1 (8) [M+Na]⁺, 478.1 (10) [M+K]⁺, 901.0 (2) [2M+Na]⁺, 917.1(3) [2M+K]⁺, 939.1 (4) [2M−H+Na+K]⁺, 945.1 (4); HRMS for C₂₃H₃₀N₅O₄[M+H]⁺ 440.2273 (calc. 440.2298); analytical HPLC (5–90% in 30 min)t_(R)=14.69 min (92.7% purity at 220 nm)

Example 5a)

Resin-bound Fmoc-1,2,3,4-tetrahydroisoquinolin-3(S)-ylacetic acid

200 mg of trityl chloride-polystyrene resin (0.18 mmol theoreticalloading) are washed in 1.5 ml of abs. DCM. A solution of 0.24 mmol ofFmoc-1,2,3,4-tetrahydroisoquinolin-3(S)-ylacetic acid and 0.6 mmol ofDIPEA in 1.5 ml of abs. DCM is subsequently added to the resin, themixture is shaken for 1.5 hours at room temperature, and 0.2 ml ofmethanol is then added. The mixture is washed with DCM (5×1.5 ml) andmethanol (3×1.5 ml) and dried.

Example 5b)

Resin-bound Fmoc-Gly-1,2,3,4-tetrahydroisoquinolin-3-ylacetic acid

0.072 mmol of A is washed with DMF (1×2 ml). The compound issubsequently deprotected twice using 20% of piperidine in DMF (2×2 ml),firstly for 5 minutes and then for 15 minutes, and washed with DMF (6×2ml). An approximately 0.1M solution of 2.5 equivalents (based on theresin loading, 0.18 mmol) of Fmoc-glycine, 2.4 equivalents (0.17 mmol)of HATU and 30 equivalents (2.16 mmol) of sym-collidine in dry DMF isadded to the resin-bound free amine, and the mixture is shaken at roomtemperature for 90 minutes. The reaction is terminated by washing in DMF(6×2 ml).

Example 5c)

3-Guanidinobenzoylglycyl-1,2,3,4-tetrahydroisoquinolin-3-ylacetic acid

0.036 mmol of B is washed with DMF (1×1 ml). The compound issubsequently deprotected twice using 20% of piperidine in DMF (2×1 ml),firstly for 5 minutes and then for 15 minutes, and washed with DMF (6×1ml). An approximately 0.1 M solution of 2.5 equivalents (based on theresin loading, 0.09 mmol) of Fmoc-3-aminobenzoic acid, 2.4 equivalents(0.086) of HATU and 30 equivalents (1.08) of sym-collidine in dry DMF isadded to the resin-bound free amine, and the mixture is shaken at roomtemperature for 90 minutes. The mixture is washed with DMF (6×1 ml) anddeprotected as described.

The resin is subsequently washed with anhydrous chloroform (3×1 ml), asolution of 0.36 mmol of N,N′-bis-BOC-1-guanylpyrazole in 0.4 ml ofanhydrous chloroform is added, and the mixture is reacted in a heatableshaker at 50° C. After 20 hours, the resin is washed with DCM (6×1 ml).For removal from the resin with simultaneous removal of BOC, the resinis shaken with a 4.75:4.75:0.5 mixture of DCM, TFA and TIPS (3×1 ml),once for 1.5 hours, once for 30 minutes and once for 3 minutes, andfiltered off The combined filtrates are evaporated, and the residue islyophilised from tert-butanol/water. Purification using preparative HPLCgives 3-guanidinobenzoylglycyl-1,2,3,4-tetrahydroisoquinolin-3-ylaceticacid, trifluoroacetate.

RT=12.3 (10→90% ACN, 30 min)

MS (ESI): m/e=410.2 ([M+H]⁺). ¹H-NMR (1.3:1 ratio of the rotationalisomers, * smaller rotational isomer signals, 500 MHz, DMSO-d₆) δ=12.39(br. s, 1H, COOH), 9.95 (s, 1H, NH—^(Ar)C), 8.67 (t, J=5.4 Hz, 1H,NH—^(Gly)CH₂), 8.63* (t, J=5.4 Hz, 1H, NH—^(Gly)CH₂), 7.78 (d, J=7.8 Hz,1H, ^(Ar)C⁶—H), 7.72 (s, 1H, ^(Ar)C²—H), 7.58 (s, 4H, ^(Gua)N₂H₄), 7.53(t, J=7.8 Hz, 1H, ^(Ar)C⁵—H), 7.39 (d, J=7.8 Hz, 1H, ^(Ar)C⁴—H),7.17–7.25 (m, 4H, ^(Thiqu)C^(5,6,7,8)—H), 5.11 (d, J=17.9 Hz, 1H,^(Thiqu)C¹—H₂), 5.01–5.02* (m, 1H, ^(Thiqu)C³—H), 4.82* (d, J=16.2 Hz,1H, ^(Thiqu)C¹—H₂), 4.73–4.75 (m, 1H, ^(Thiqu)C³—H), 4.55* (d, J=16.2Hz, 1H, ^(Thiqu)C¹—H₂), 4.44 (dd, J=16.4 Hz, J=5.4 Hz, 1H, ^(Gly)CH₂),4.19–4.29 (m, 3H, ^(Gly)CH₂), 4.10 (d, J=17.9 Hz, 1H, ^(Thiqu)C¹—H₂),3.15 (dd, J=16.4 Hz, J=5.0 Hz, 1H CH₂CO₂H), 2.98* (dd, J=15.8 Hz, J=5.2Hz, 1H, CH₂CO₂H), 2.74–2.79 (m, 2H, CH₂CO₂H), 2.41–2.51, 2.33–2.37,2.16–2.21 (m, 4H, ^(Thiqu)C⁴—H₂).

Example 6

5-(4-Methylpyridin-2-ylamino)pentanoylglycyl-1,2,3,4-tetrahydroisoquinolin-3-ylaceticacid

0.036 mmol of B (obtainable as described in Example. 5b)) is washed withDMF (1×1 ml). The compound is subsequently deprotected twice using 20%of piperidine in DMF (2×1 ml), firstly for 5 minutes and then for 15minutes, and washed with DMF (6×1 ml). The resin-bound free amine isshaken overnight at room temperature with an approximately 0.1 Msolution of 2.5 equivalents (0.09 mmol) of5-(N-(4-methylpyridin-2-yl)amino-pentanoic acid, 2.4 equivalents (0.086mmol) of HATU and 30 equivalents (1.08 mmol) of collidine in absoluteDMF. The mixture is washed with DMF and DCM. For removal from the solidphase, the washed resin is shaken with 1 ml of a mixture ofDCM/trifluoroethanol/acetic acid (31/1), firstly for 90 minutes, thenfor 30 minutes and finally for 1 minute. Removal of the solvent andpurification using preparative HPLC gives5-(4-methylpyridin-2-ylamino)pentanoylglycyl-1,2,3,4-tetrahydroisoquinolin-3-ylaceticacid, trifluoroacetate.

RT=13.3 (10→90% ACN, 30 min)

MS (ESI): m/e=439.3 ([M+H]⁺).

¹H-NMR (1.3:1 ratio of the rotational isomers, * smaller rotationalisomer signals, 500 MHz, DMSO-d₆) δ=12.35 (br. s, 1H, COOH), 8.46 (br.s, 1H, NH—CH₂), 7.93–7.97 (m, 1H, NH—^(Gly)CH₂), 7.78 (d, J=6.5 Hz, 1H,^(Pyr)C⁶—H), 7.14–7.22 (m, 4H, ^(Thiqu)C^(5,6,7,8)—H), 6.81 (s, 1H,^(Pyr)C³—H), 6.69 (d, J=6.5 Hz, 1H, ^(Pyr)C⁵—H), 5.08 (d, J=17.9 Hz, 1H,^(Thiqu)C¹—H₂), 4.97–5.01* (m, 1H, ^(Thiqu)C³—H), 4.71* (d, J=16.2 Hz,1H, ^(Thiqu)C¹—H₂4.59–4.63 (m, 1H, ^(Thiqu)C³—H), 4.47* (d, J=16.2 Hz,1H, ^(Thiqu)C¹—H₂) (dd, J=16.7 Hz, J=5.5 Hz, 1H, ^(Gly)CH₂), 4.04–4.08(m, 3H, ^(Gly)CH₂, ^(Thiqu)C¹—H₂), 3.97* (dd, J=16.9 Hz, J=5.4 Hz, 1H,^(Gly)CH₂), 3.27 (m, 2H, NH—CH₂), 3.11 (dd, J=16.2 Hz, J=5.3 Hz, 1H,CH₂CO₂H), 2.95* (dd, J=15.7 Hz, J=5.4 Hz, 1H, CH₂CO₂H), 2.62–2.77 (m,2H, CH₂CO₂H), 2.34–2.44 (m, 3H, ^(Thiqu)C⁴—H₂) 2.31 (s, 3H, CH₃),2.12–2.21 (m, 3H, ^(Thiqu)C⁴—H₄, CH₂—CH₂—CO), 1.58 (s, 4H,(CH₂)₂—CH₂—CO).

The other compounds of the formula I, in particular the compounds of theformulae I1 to I36, can be obtained analogously using the correspondingprecursors.

The examples below relate to pharmaceutical preparations:

Example A Injection Vials

A solution of 100 g of an active ingredient of the formula I and 5 g ofdisodium hydrogenphosphate in 3 l of bidistilled water is adjusted to pH6.5 using 2N hydrochloric acid, sterile filtered, transferred intoinjection vials, lyophilised under sterile conditions and sealed understerile conditions. Each injection vial contains 5 mg of activeingredient.

Example B Suppositories

A mixture of 20 g of an active ingredient of the formula I is meltedwith 100 g of soya lecithin and 1400 g of cocoa butter, poured intomoulds and allowed to cool. Each suppository contains 20 mg of activeingredient.

Example C Solution

A solution is prepared from 1 g of an active ingredient of the formulaI, 9.38 g of NaH₂PO₄.2 H₂O, 28.48 g of Na₂HPO₄.12 H₂O and 0.1 g ofbenzalkonium chloride in 940 ml of bidistilled water. The pH is adjustedto 6.8, and the solution is made up to 1 l and sterilised byirradiation. This solution can be used in the form of eye drops:

Example D Ointment

500 mg of an active ingredient of the formula I are mixed with 99.5 g ofVaseline under aseptic conditions.

Example E Tablets

A mixture of 1 kg of active ingredient of the formula I, 4 kg oflactose, 1.2 kg of potato starch, 0.2 kg of talc and 0.1 kg of magnesiumstearate is pressed to give tablets in a conventional manner in such away that each tablet contains 10 mg of active ingredient.

Example F Coated Tablets

Tablets are pressed analogously to Example E and subsequently coated ina conventional manner with a coating of sucrose, potato starch, talc,tragacanth and dye.

Example G Capsules

2 kg of active ingredient of the formula I are introduced into hardgelatine capsules in a conventional manner in such a way that eachcapsule contains 20 mg of the active ingredient.

Example H Ampoules

A solution of 1 kg of active ingredient of the formula I in 60 ofbidistilled water is sterile filtered, transferred into ampoules,lyophilised under sterile conditions and sealed under sterileconditions. Each ampoule contains 10 mg of active ingredient.

Example I Inhalation Spray

14 g of active ingredient of the formula I are dissolved in 10 l ofisotonic NaCl solution, and the solution is transferred intocommercially available spray containers with a pump mechanism. Thesolution can be sprayed into the mouth or nose. One spray shot (about0.1 ml) corresponds to a dose of about 0.14 mg.

1. A compound of the formula I:

in which X is H, —C(═NR³)—NHR⁴ or Het,

Y is —(CH₂)_(m)-, Z is NH or CH₂, R¹ and R⁵ are each, independently ofone another, H, A, OH, OA, arylalkyl, Hal, —CO—A, CN, NO₂, NHR³, COOA,COOH, SO₂A, CF₃ or OCF₃, R² is in each case, independently of theothers, H or A, R³ and R⁴ are each, independently of one another, H, A,—CO—A, NO₂ or CN, A is alkyl having 1–6 carbon atoms, m is 0, 1, 2, 3,4, 5 or 6, n and p are, independently of one another, 1, 2 or 3; or aphysiologically acceptable salt or solvate thereof.
 2. A compound of theformula I of claim 1, wherein A is methyl, ethyl, isopropyl, n-propyl,n-butyl, isobutyl, sec-butyl or tert-butyl.
 3. A compound of the formulaI of claim 1, wherein Het is 4-methylpyridin-2-yl, pyridin-2-yl,pyrimidin-2-yl, imidazol-2-yl, benzimidazol-2-yl or a hydrogenated groupthereof.
 4. A compound of the formula I of claim 1, wherein R¹ and R⁵,independently of one another, are H, A, CN, NO₂, Hal or —COA.
 5. Acompound of the formula I of claim 1, wherein R² H or A.
 6. A compoundof the formula I of claim 1, wherein R³ and R⁴, independently of oneanother, are H or —COA.
 7. A compound of the formula I of claim 1,wherein X is H, —C(═NH)—NH₂, —C(═N-methyl)-NH₂, 4-methylpyridin-2-yl,pyridin-2-yl, pyrimidin-2-yl, imidazol-2-yl, benzimidazol-2-yl or ahydrogenated group thereof.
 8. A compound of the formula I of claim 1,wherein Y is —(CH₂)_(m)- or


9. A compound of the formula I of claim 1, wherein n and p,independently of one another, are 1 or
 2. 10. A compound of the formulaI of claim 1, wherein m is 0, 2 or
 4. 11. A compound of the formula I ofclaim 1, which is of one of the following formulae I1 to I36:


12. A method for preparing a compound of the formula I of claim 1, whichcomprises: a) reacting a compound of the formula II

in which Z, R¹ and n are as defined above, and W is a conventionalprotecting group or a solid phase used in peptide chemistry, with acompound of the formula III

in which Y is as defined above, and Q is a suitable protecting group orHet, in the presence of a condensing agent, removing at least oneprotecting group and/or the solid phase, and, optionally, where Q asprotecting group is removed, reacting the resultant product with aguanyl compound and, optionally, removing any remaining protectinggroups and/or the solid phase, or b) liberating a compound of theformula I from a functional derivative of a compound of the formula I bytreatment with a solvolyzing or hydrogenolyzing agent, and/or c)converting a basic or acidic compound of the formula I into one of itssalts by treatment with an acid or base.
 13. A pharmaceuticalcomposition comprising a compound of the formula I of claim 1 or aphysiologically acceptable salt or solvate thereof and at least onesolid, liquid and/or semi-liquid excipient or adjuvant.
 14. Acomposition of claim 13 comprising an amount of the compound of theformula I effective to provide integrin inhibitor activity.
 15. A methodfor preparing a composition of claim 13 which comprises formulating acompound of the formula I with at least one solid, liquid and/orsemi-liquid excipient or adjuvant.
 16. A method for treating thromboses,cardiac infarction, coronary heart disease, arteriosclerosis,inflammation, tumours, osteoporosis, an infection or restenosis afterangioplasty, which comprises administering to a patient in need thereofa compound of formula I of claim 1 or a physiologically acceptable saltor solvate thereof.